De-differentiation protocols are useful for creating pluripotent cells from adult connective tissue for experimentation, amplification and clinical applications. Current de-differentiation protocols to achieve pluripotent cells are inefficient and limited. Furthermore, the use of growth factors and manipulation of culture conditions have been utilized to increase proliferation rates of cell cultures with limited success.
Typically, in culturing fibroblasts, primary fibroblasts are used. One major limitation in using primary cells for the study of disease is their limited in vitro lifespan and contributes to why primary adult derived human cell lines for are used infrequently in genetic research. During the process of DNA transfection, and subsequent cell amplification, primary cells reach their in vitro proliferation potential making extended studies impossible. Thus, a need exists for increasing the life span of cultured connective tissue for extended studies on successfully genetically modified cells.
Additionally, utilizing adult fibroblast and developing cultures specific to individuals or animals would be advantageous in clinical applications for these individuals. Thus, a need exists for generating de-differentiated connective tissue cells from an individual.